1,721,433 research outputs found
Regulation of adenovirus replication by miR-199 confers a selective oncolytic activity in hepatocellular carcinoma
No full textOncolytic virotherapy represents a growing field of experimental cancer therapy. For safe and effective virotherapy, restricted tissue expression and replication of the virus is desirable. Various methods have been developed to achieve such restricted expression. They included the engineering of viral genomes through the insertion of tissue-specific promoters or genes encoding for tissue specific binding proteins. Here, we employed a new approach based on the use of microRNAs (miRNAs) to achieve tumor-specific viral expression and replication. miRNAs are approximately 22-nucleotide (nt)- long non-coding RNAs that are able to bind the 3’ untranslated regions (UTRs) of homologous target mRNAs and causing either their degradation or translation inhibition. Since miRNA are differentially expressed in cancer versus normal cells, it is theoretically possible to make virus expression restricted to cancer cells in a miRNA-dependent manner.
Several studies have shown that miR-199 is significantly down-regulated in primary hepatocellular carcinoma (HCC) tissue and HCC cell lines. With this notion in mind, we developed a conditionally replication-competent oncolytic adenovirus, Ad-199T, by introducing four copies of miR-199 target sites within the 3′ UTR of the E1A gene, which is essential for adenovirus replication.
In vitro studies of the properties of Ad-199T virus revealed that E1A expression was indeed tightly regulated both at RNA and protein levels depending upon the expression of miR-199. Consequently, Ad-199T could replicate in the HCC derived cells HepG2, negative for miR-199 expression, while its replication was strictly controlled in HepG2-199 cells, which were engineered to express high level of miR-199. A replication-competent miRNA independent Ad-Control was also generated,. Thus, these in vitro studies proved that cytotoxicity of Ad-199T was effective in HCC derived cells, which lacks expression of miR-199, and could be successfully controlled in cells that express miR-199 at high level.
To assess in vivo properties of Ad-199T, we tested an orthotopic tumor model. HepG2 cells were implanted in the liver of newborn B6D2 mice. The cells could survive at least one week in this environment, enough for testing in vivo properties of Ad-199T. These studies revealed that intrahepatic delivery of Ad-199T led to virus replication in HepG2 derived xenograft tumors and a faster removal of cancer cells. Conversely, Ad-199T replication was not detected in normal, miR-199 positive, liver parenchyma.
These results demonstrate that Ad-199T is a conditionally replicative adenovirus (CRAd) miR-199 dependent, with antitumor activity in vivo. This system allows replication of the oncolytic virus in HCC cells and, at the same time, tightly control replication in normal liver tissues, thus avoiding or reducing hepatotoxicity
Generation and validation of MIR-142 knock out mice
No full text© 2015 Shrestha et al.This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. microRNA-142 (miR-142) is an important regulator of many biological processes and associated signaling pathways during embryonic development, homeostasis and disease. The miR-142 hairpin gives rise to the "guide strand" miR-142-3p and the sister "passenger" strand miR-142-5p. miR-142-3p has been shown to play critical, non-redundant functions in the development of the hematopoietic lineage. We have recently reported that miR-142-3p is critical for the control of Wnt signaling in the mesenchyme of the developing lung. miR-142-5p has been proposed to control adaptive growth in cardiomyocytes postnatally and its increase is associated with extensive apoptosis and cardiac dysfunction in a murine heart failure model. Using homologous recombination, we now report the generation and validation of miR-142-null mice. miR-142-null mice show a significant decrease in th expression levels of both the 3p and 5p isoforms. The expression of Bzrap1, a gene immediately flanking miR-142 is not altered while the expression of a long non-coding RNA embedded within the miR-142 gene is decreased. miR-142-null newborn pups appear normal and are normally represented indicating absence of embryonic lethality. At embryonic day 18.5, miR-142-null lungs display increased Wnt signaling associated with the up-regulation of Apc and p300, two previously reported targets of miR-142-3p and -5p, respectively. Adult miR-142-null animals display impaired hematopoietic lineage formation identical to previously reported miR-142 gene trap knockdown mice. We report, for the first time, the homologous recombination-based miR-142-null mice that will be useful for the scientific community working on the diverse biological functions of miR-142
Generation and validation of MIR-142 knock out mice
Get PDF© 2015 Shrestha et al.This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. microRNA-142 (miR-142) is an important regulator of many biological processes and associated signaling pathways during embryonic development, homeostasis and disease. The miR-142 hairpin gives rise to the "guide strand" miR-142-3p and the sister "passenger" strand miR-142-5p. miR-142-3p has been shown to play critical, non-redundant functions in the development of the hematopoietic lineage. We have recently reported that miR-142-3p is critical for the control of Wnt signaling in the mesenchyme of the developing lung. miR-142-5p has been proposed to control adaptive growth in cardiomyocytes postnatally and its increase is associated with extensive apoptosis and cardiac dysfunction in a murine heart failure model. Using homologous recombination, we now report the generation and validation of miR-142-null mice. miR-142-null mice show a significant decrease in th expression levels of both the 3p and 5p isoforms. The expression of Bzrap1, a gene immediately flanking miR-142 is not altered while the expression of a long non-coding RNA embedded within the miR-142 gene is decreased. miR-142-null newborn pups appear normal and are normally represented indicating absence of embryonic lethality. At embryonic day 18.5, miR-142-null lungs display increased Wnt signaling associated with the up-regulation of Apc and p300, two previously reported targets of miR-142-3p and -5p, respectively. Adult miR-142-null animals display impaired hematopoietic lineage formation identical to previously reported miR-142 gene trap knockdown mice. We report, for the first time, the homologous recombination-based miR-142-null mice that will be useful for the scientific community working on the diverse biological functions of miR-142
Edna O’Brien as a postmodernist novelist.
Get PDFEdna O’Brien is one of the most prolific and versatile writers of the twentieth century. She made her debut in 1960 with The Country Girls and since then has authored many novels, short stories, plays, poems, screenplays, children’s fiction and non-fictional prose.
In spite of being a prolific writer, she is little known outside Ireland, the United Kingdom and the United States of America. Though being a recipient of many literary awards, she has not been recognized worldwide. By and large, her works have suffered neglect from literary critics and scholars, and whatsoever has been written has been written from a limited and orthodox point of view. There are two main reasons for this neglect by the literary critical establishment. One is that critics focused more attention on the so-called ‘autobiographical’ nature of her works. This tendency proved very detrimental for the author. Amanda Greenwood writes, “The reductive tendency to identify the author with her female protagonists has been largely instrumental in her relegation to ‘popular fiction’.” The other reason is that her own dandyish personality, flamboyant nature and nationality, which made her to be stereotyped as Celtic and wild, attracted greater attention than any serious critical debate on her works. Claudia Pattison commented in a review that “with her flame-red hair, milky skin and mesmerising green-flecked eyes, [O’Brien] is a bewitching Celtic beauty.” Her beauty and persona led critics to doubt her writing ability. David Goodwin agreed that “because Edna is a more glamorous woman sometimes people don’t take her so seriously.”3 Edna O’Brien was herself well aware of this fact. She once remarked that “if you happen to have your hair done, well, then you can’t be a ‘serious’ writer.”4 Rebecca Pelan also points out that it was O’Brien’s public persona which led to “the relegation of [her] writing to the realm of popular fiction, a … category which allows the content of her writing to go virtually unnoticed.”However, she always insisted on being a serious writer. She once said to a reviewer, “I am a serious writer. Take more notice of the books than how i look.”Digital copy of Ph.D thesis.University of Kashmir
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Dispelling the Myths Behind First-author Citation Counts
Get PDFWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
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